Potential long-term benefits of acute hypothermia after spinal cord injury: assessments with somatosensory-evoked potentials.

OBJECTIVE: Neuroprotection by hypothermia has been an important research topic over last two decades. In animal models of spinal cord injury, the primary focus has been assessing the effects of hypothermia on behavioral and histologic outcomes. Although a few studies have investigated electrophysiological changes in descending motor pathways with motor-evoked potentials recorded during cooling, we report here hypothermia induced increased electrical conduction in the ascending spinal cord pathways with somatosensory-evoked potentials in injured rats. In our experiments, these effects lasted long after the acute hypothermia and were accompanied by potential long-term improvements in motor movement. DESIGN: Laboratory investigation. SETTING: University medical school. SUBJECTS: Twenty-one female Lewis rats. INTERVENTIONS: Hypothermia. MEASUREMENTS AND MAIN RESULTS: All animals underwent spinal cord contusion with the NYU-Impactor by a 12.5-mm weight drop at thoracic vertebra T8. A group (n = 10) was randomly assigned for a systemic 2-hr hypothermia episode (32 ± 0.5°C) initiated approximately 2.0 hrs postinjury. Eleven rats were controls with postinjury temperature maintained at 37 ± 0.5°C for 2 hrs. The two groups underwent preinjury, weekly postinjury (up to 4 wks) somatosensory-evoked potential recordings and standard motor behavioral tests (BBB). Three randomly selected rats from each group were euthanized for histologic analysis at postinjury day 3 and day 28. Compared with controls, the hypothermia group showed significantly higher postinjury somatosensory-evoked potential amplitudes with longer latencies. The BBB scores were also higher immediately after injury and 4 wks later in the hypothermia group. Importantly, specific changes in the Basso, Beattie, Bresnahan scores in the hypothermia group (not seen in controls) indicated regained functions critical for motor control. Histologic evaluations showed more tissue preservation in the hypothermia group. CONCLUSIONS: After spinal cord injury, early systemic hypothermia provided significant neuroprotection weeks after injury through improved sensory electrophysiological signals in rats. This was accompanied by higher motor behavioral scores and more spared tissue in acute and postacute periods after injury.

The Do-It-Yourself Electrocardiogram.

Hands-on labs are a critical component of biomedical engineering undergraduate education. Due to both the pandemic and the growing interest in online education, we developed a Do-it-yourself Electrocardiogram (DIY EKG) project. The Arduino-based DIY EKG kit instructed students how to build a circuit to obtain their own EKG and then analyze their EKG data using Matlab. Despite the obstacles of virtually trouble-shooting, 85.4% of students (n = 103) were able to obtain their own EKG at home. We have provided the labelled circuit drawings, step-by-step instructions, Matlab files, and results in this paper. Survey results indicate that 89% of students felt the DIY EKG project was a "challenging yet fulfilling experience." Supplementary Information: The online version of this article contains supplementary material available 10.1007/s43683-021-00061-0.

Human glial-restricted progenitors survive, proliferate, and preserve electrophysiological function in rats with focal inflammatory spinal cord demyelination.

Transplantation of glial progenitor cells results in transplant-derived myelination and improved function in rodents with genetic dysmyelination or chemical demyelination. However, glial cell transplantation in adult CNS inflammatory demyelinating models has not been well studied. Here we transplanted human glial-restricted progenitor (hGRP) cells into the spinal cord of adult rats with inflammatory demyelination, and monitored cell fate in chemically immunosuppressed animals. We found that hGRPs migrate extensively, expand within inflammatory spinal cord lesions, do not form tumors, and adopt a mature glial phenotype, albeit at a low rate. Human GRP-transplanted rats, but not controls, exhibited preserved electrophysiological conduction across the spinal cord, though no differences in behavioral improvement were noted between the two groups. Although these hGRPs myelinated extensively after implantation into neonatal shiverer mouse brain, only marginal remyelination was observed in the inflammatory spinal cord demyelination model. The low rate of transplant-derived myelination in adult rat spinal cord may reflect host age, species, transplant environment/location, and/or immune suppression regime differences. We conclude that hGRPs have the capacity to myelinate dysmyelinated neonatal rodent brain and preserve conduction in the inflammatory demyelinated adult rodent spinal cord. The latter benefit is likely dependent on trophic support and suggests further exploration of potential of glial progenitors in animal models of chronic inflammatory demyelination.

Quantitative assessment of somatosensory-evoked potentials after cardiac arrest in rats: prognostication of functional outcomes.

OBJECTIVE: High incidence of poor neurologic sequelae after resuscitation from cardiac arrest underscores the need for objective electrophysiological markers for assessment and prognosis. This study aims to develop a novel marker based on somatosensory evoked potentials (SSEPs). Normal SSEPs involve thalamocortical circuits suggested to play a role in arousal. Due to the vulnerability of these circuits to hypoxic-ischemic insults, we hypothesize that quantitative SSEP markers may indicate future neurologic status. DESIGN: Laboratory investigation. SETTING: University Medical School and Animal Research Facility. SUBJECTS: : Sixteen adult male Wistar rats. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: SSEPs were recorded during baseline, during the first 4 hrs, and at 24, 48, and 72 hrs postasphyxia from animals subjected to asphyxia-induced cardiac arrest for 7 or 9 mins (n = 8/group). Functional evaluation was performed using the Neurologic Deficit Score (NDS). For quantitative analysis, the phase space representation of the SSEPs-a plot of the signal vs. its slope-was used to compute the phase space area bounded by the waveforms recorded after injury and recovery. Phase space areas during the first 85-190 mins postasphyxia were significantly different between rats with good (72 hr NDS >or=50) and poor (72 hr NDS <50) outcomes (p = .02). Phase space area not only had a high outcome prediction accuracy (80-93%, p < .05) during 85-190 mins postasphyxia but also offered 78% sensitivity to good outcomes without compromising specificity (83-100%). A very early peak of SSEPs that precedes the primary somatosensory response was found to have a modest correlation with the 72 hr NDS subscores for thalamic and brainstem function (p = .066) and not with sensory-motor function (p = .30). CONCLUSIONS: Phase space area, a quantitative measure of the entire SSEP morphology, was shown to robustly track neurologic recovery after cardiac arrest. SSEPs are among the most reliable predictors of poor outcome after cardiac arrest; however, phase space area values early after resuscitation can enhance the ability to prognosticate not only poor but also good long-term neurologic outcomes.

Evoked potential and behavioral outcomes for experimental autoimmune encephalomyelitis in Lewis rats.

A reliable outcome measurement is needed to assess the effects of experimental lesions in the rat spinal cord as well as to assess the benefits of therapies designed to modulate them. The Basso, Beattie, and Bresnahan (BBB) behavioral scores can be indicative of the functionality in motor pathways. However, since lesions are often induced in the more accessible dorsal parts associated with the sensory pathways, the BBB scores may not be ideal measure of the disability. We propose somatosensory evoked potential (SEP) as a complementary measure to assess the integrity of sensory pathways. We used the focal experimental autoimmune encephalomyelitis (EAE) model, in which focal demyelinating lesions were induced by injecting cytokine-ethidium bromide into dorsal white matter after MOG-IFA immunization. Both the SEP and BBB measures reflected injury; however, the SEP was uniformly and consistently altered after the injury whereas the BBB varied widely. The results suggest that the SEP measures are more sensitive and reliable markers of focal spinal cord demyelination compared to the behavioral measures like the BBB score.

Detection of repolarization alternans with an implantable cardioverter defibrillator lead in a porcine model.

Mechanistic links have been suggested between repolarization alternans (RPA) and the onset of ventricular tachycardia (VT) and/or fibrillation. Endocardial detection of RPA may, therefore, be an important step in future device-based treatments of arrhythmias. Here, we investigate if RPA could be detected during acute ischemia using an implantable cardioverter defibrillator (ICD) lead (tip to distal coil) located in the right ventricular apex. In 18 pigs, the right coronary (n = 10) or left anterior descending coronary (n = 8) artery was occluded for 10 min using a balloon catheter, followed by reperfusion for 30 min, and re-occlusion for 30 min. RPA magnitude, computed using the modified moving average (MMA) method, showed a sharp increase in all 18 animals, from a mean baseline level of 1.9 +/- 1.3 mV to 3.0 +/- 1.3 mV during first occlusion (p < 0.001). RPA magnitude showed a prominent increase in 10 animals during re-occlusion, from a mean baseline level of 1.7 +/- 1.0 mV to 3.3 +/- 1.5 mV (p < 0.001). The protocol was terminated during the first two stages of occlusion and reperfusion for the remaining 8 animals due to the occurrence of ventricular fibrillation (VF). These results confirm that RPA increases under ischemic conditions and that it is possible to detect and track RPA dynamics with an ICD lead that is positioned in a clinically realistic location. Such an approach may be useful in formulating improved arrhythmia detection and control algorithms.

Dynamics of stick-slip in peeling of an adhesive tape.

We investigate the dynamics of peeling of an adhesive tape subjected to a constant pull speed. We derive the equations of motion for the angular speed of the roller tape, the peel angle and the pull force used in earlier investigations using a Lagrangian. Due to the constraint between the pull force, peel angle and the peel force, it falls into the category of differential-algebraic equations requiring an appropriate algorithm for its numerical solution. Using such a scheme, we show that stick-slip jumps emerge in a purely dynamical manner. Our detailed numerical study shows that these set of equations exhibit rich dynamics hitherto not reported. In particular, our analysis shows that inertia has considerable influence on the nature of the dynamics. Following studies in the Portevin-Le Chatelier effect, we suggest a phenomenological peel force function which includes the influence of the pull speed. This reproduces the decreasing nature of the rupture force with the pull speed observed in experiments. This rich dynamics is made transparent by using a set of approximations valid in different regimes of the parameter space. The approximate solutions capture major features of the exact numerical solutions and also produce reasonably accurate values for the various quantities of interest.

Enhancement of bilateral cortical somatosensory evoked potentials to intact forelimb stimulation following thoracic contusion spinal cord injury in rats.

The adult central nervous system is capable of significant reorganization and adaptation following neurotrauma. After a thoracic contusive spinal cord injury (SCI) neuropathways that innervate the cord below the epicenter of injury are damaged, with minimal prospects for functional recovery. In contrast, pathways above the site of injury remain intact and may undergo adaptive changes in response to injury. We used cortical somatosensory evoked potentials (SSEPs) to evaluate changes in intact forelimb pathways. Rats received a midline contusion SCI, unilateral contusion SCI, or laminectomy with no contusion at the T8 level and were monitored for 28 days post-injury. In the midline injury group, SSEPs recorded from the contralateral forelimb region of the primary somatosensory cortex were 59.7% (CI 34.7%, 84.8%; c(2) = 21.9; dof = 1; p = 2.9 ×10(-6)) greater than the laminectomy group; SSEPs from the ipsilateral somatosensory cortex were 47.6% (CI 18.3%, 77%; c(2) = 10.1; dof = 1; p = 0.001) greater. Activation of the ipsilateral somatosensory cortex was further supported by BOLD-fMRI, which showed increased oxygenation at the ipsilateral hemisphere at day seven post-injury. In the unilateral injury group, ipsilesional side was compared to the contralesional side. SSEPs on day 14 (148%; CI 111%, 185%) and day 21 (137%; CI 110%, 163%) for ipsilesional forelimb stimulation were significantly increased over baseline (100%). SSEPs recorded from the hindlimb sensory cortex upon ipsilesional stimulation were 33.9% (CI 14.3%, 53.4%; c(2) = 11.6; dof = 1; p = 0.0007) greater than contralesional stimulation. Therefore, these results demonstrate the ability of SSEPs to detect significant enhancements in the activation of forelimb sensory pathways following both midline and unilateral contusive SCI at T8. Reorganization of forelimb pathways may occur after thoracic SCI, which SSEPs can monitor to aid the development of future therapies.

Effect of hypothermia on cortical and thalamic signals in anesthetized rats.

Beneficial effects of hypothermia on subjects with neuro-pathologies have been well demonstrated in both animal studies and clinical trials. Although it is known that temperature significantly impacts neurological injuries, the underlying mechanism remains unclear. We studied the effect of temperature modulation on neural signals in the cortex and the thalamus in uninjured brains of anesthetized rats. Six rats were divided into a hypothermic (32 to 34 °C, n=3) and a hyperthermic group (38.5 to 39.5 °C, n=3). EEG, and extracellular signals from somatosensory cortex and the ventral posterolateral nucleus of thalamus were recorded at different temperature phases (normothermia (36.5 to 37.5 °C) and hypothermia or hyperthermia). During hypothermia, similar burst suppression (BS) patterns were observed in cortical and thalamic signals as in EEG, but thalamic activity was not completely under suppression when both EEG and cortical signals were electrically silent. In addition, our results showed that hypothermia significantly increased the burst suppression ratio (BSR) in EEG, cortical and thalamic signals by 3.42, 3.25, 7.29 times respectively (P<0.01), and prolonged the latency of neuronal response in cortex to median nerve stimulation from 9 ms to 16 ms (P<0.01). Furthermore, during normothermia, the correlation coefficient between thalamic and cortical signals was 0.35±0.02 while during hypothermia, it decreased to 0.16±0.03 with statistical significance (P<0.01). These results can potentially assist in better understanding the effects of hypothermia.

Assessing thalamocortical functional connectivity with Granger causality.

Assessment of network connectivity across multiple brain regions is critical to understanding the mechanisms underlying various neurological disorders. Conventional methods for assessing dynamic interactions include cross-correlation and coherence analysis. However, these methods do not reveal the direction of information flow, which is important for studying the highly directional neurological system. Granger causality (GC) analysis can characterize the directional influences between two systems. We tested GC analysis for its capability to capture directional interactions within both simulated and in vivo neural networks. The simulated networks consisted of Hindmarsh-Rose neurons; GC analysis was used to estimate the causal influences between two model networks. Our analysis successfully detected asymmetrical interactions between these networks ( , t -test). Next, we characterized the relationship between the "electrical synaptic strength" in the model networks and interactions estimated by GC analysis. We demonstrated the novel application of GC to monitor interactions between thalamic and cortical neurons following ischemia induced brain injury in a rat model of cardiac arrest (CA). We observed that during the post-CA acute period the GC interactions from the thalamus to the cortex were consistently higher than those from the cortex to the thalamus ( 1.983±0.278 times higher, p = 0.021). In addition, the dynamics of GC interactions between the thalamus and the cortex were frequency dependent. Our study demonstrated the feasibility of GC to monitor the dynamics of thalamocortical interactions after a global nervous system injury such as CA-induced ischemia, and offers preferred alternative applications in characterizing other inter-regional interactions in an injured brain.

Band specific changes in thalamocortical synchrony in field potentials after cardiac arrest induced global hypoxia.

Cardiac Arrest (CA) leads to a global hypoxic-ischemic injury in the brain leading to a poor neurological outcome. Understanding the mechanisms of functional disruption in various regions of the brain may be essential for the development of improved diagnostic and therapeutic solutions. Using controlled laboratory experiment with animal models of CA, our primary focus here is on understanding the functional changes in the thalamus and the cortex, associated with the injury and acute recovery upon resuscitation. Specifically, to study the changes in thalamocortical synchrony through these periods, we acquired local field potentials (LFPs) from the ventroposterior lateral (VPL) nucleus of the thalamus and the forelimb somatosensory cortex (S1FL) in rats after asphyxial CA. Band-specific relative Hilbert phases were used to analyze synchrony between the LFPs. We observed that the CA induced global ischemia changes the local phase-relationships by introducing a phase-lag in both the thalamus and the cortex, while the synchrony between the two regions is nearly completely lost after CA.

Effect of hypothermia on the thalamocortical function in the rat model.

Neuroprotective effects of hypothermia are well documented in many injuries of the central nervous system in animal models as well as clinical studies. However, the underlying mechanisms are not fully understood. An important yet unexplored background issue is the effect of hypothermic cooling on the regional functionality of the healthy CNS. In a pilot study with the rat model, we seek to characterize the effect of moderate bodily cooling on the thalamo-cortical (T-C) function. Multiunit activity (MUA) and local field potentials (LFPs) were recorded from the thalamus (VPL nucleus) and the somatosensory cortex (S1) for normothermic, mild hypothermic and mild hyperthermic conditions in healthy rats and the thalamo-cortical dynamics was characterized with Granger Causal Interaction (GCI). The GCI indicated that the thalamic driving of the cortical activity significantly increases in strength with bodily cooling and weakens with mild heating. These results could have important implications towards understanding of hypothermia.

Electrophysiological evaluation of sensory and motor pathways after incomplete unilateral spinal cord contusion.

OBJECT: Unilateral contusions represent an increasingly popular model for studying the pathways and recovery mechanisms of spinal cord injury (SCI). Current studies rely heavily on motor behavior scoring and histological evidence to make assessments. Electrophysiology represents one way to reliably quantify the functionality of motor pathways. The authors sought to quantify the functional integrity of the bilateral motor and sensory pathways following unilateral SCI by using measurements of motor and somatosensory evoked potentials (MEPs and SSEPs, respectively). METHODS: Eighteen rats were randomly divided into 3 groups receiving a mild unilateral contusion, a mild midline contusion, or a laminectomy only (control). Contusions were induced at T-8 using a MASCIS impactor. Electrophysiological analysis, motor behavior scoring, and histological quantifications were then performed to identify relationships among pathway conductivity, motor function, and tissue preservation. RESULTS: Hindlimb MEPs ipsilateral to the injury showed recovery by Day 28 after injury and corresponded to approximately 61% of spared corticospinal tract (CST) tissue. In contrast, MEPs of the midline-injured group did not recover, and correspondingly > 90% of the CST tissue was damaged. Somatosensory evoked potentials showed only a moderate reduction in amplitude, with no difference in latency for the pathways ipsilateral to injury. Furthermore, these SSEPs were significantly better than those of the midline-injured rats for the same amount of white matter damage. CONCLUSIONS: Motor evoked potential recovery corresponded to the amount of spared CST in unilateral and midline injuries, but motor behavior consistently recovered independent of MEPs. These data support the idea that spared contralateral pathways aid in reducing the functional deficits of injured ipsilateral pathways and further support the idea of CNS plasticity.

Plasticity associated changes in cortical somatosensory evoked potentials following spinal cord injury in rats.

Spinal cord injury (SCI) causes a number of physiological and neurological changes resulting in loss of sensorimotor function. Recent work has shown that the central nervous system is capable of plastic behaviors post-injury, including axonal regrowth and cortical remapping. Functional integrity of afferent sensory pathways can be quantified using cortical somatosensory evoked potentials (SSEPs) recorded upon peripheral limb stimulation. We implanted 15 rats with transcranial screw electrodes and recorded SSEPs from cortical regions corresponding to each limb before and after a mild or moderate contusion injury. We report a post-injury increase in the mean amplitude of cortical SSEPs upon forelimb stimulation. SSEP amplitudes for mild and moderate SCI groups increased by 183% ± 95% and 107% ± 38% over baseline, respectively, while hindlimb SSEPs decreased by 58% ± 14% and 79% ± 4%. In addition, we report increased SSEP amplitude measured from the anatomically adjacent hindlimb region upon forelimb stimulation (increase of 90% ± 19%). Our results show that previously allocated hindlimb cortical regions are now activated by forelimb stimulation, suggesting an expansion in the area of cortical forelimb representation into hindlimb regions after an injury. This result is indicative of adaptive plasticity in undamaged areas of the CNS following SCI.

Characterization of neurologic injury using novel morphological analysis of Somatosensory Evoked Potentials.

This paper describes an innovative, easy-to-interpret, clinically translatable tool for analysis of Somatosensory Evoked Potentials (SSEPs). Unlike traditional analysis, which involves peak-to-peak amplitude and latency calculation, this method, phase space analysis, analyzes the overall morphology of the SSEP, and includes greater information. The SSEP is plotted in phase space (x-dot vs. x), which leads to an approximately spiral curve. The area swept out by this curve is termed the Phase Space Area (PSA). As PSA calculation involves numerical differentiation, we present a comparison of two different approaches to combat noise amplification: finite-window smoothing, and total variation regularization (TVR) of the numerical derivative. These methods are applied to simulated SSEPs. The efficacy of these methods in performing noise-reduction is assessed and compared with ensemble averaging. While TVR gives a reasonably robust approximation of the derivative, Gaussian smoothing of the derivative offers the best trade-off between the number of signal sweeps required to be averaged, close approximation of the SSEP derivative, and optimal estimation of the PSA. We validate this method by analyzing non-characteristic SSEPs that have indistinguishable peaks as is frequently seen in cases of underlying neurologic injury such as hypoxic-ischemic encephalopathy.

Multi-limb acquisition of motor evoked potentials and its application in spinal cord injury.

The motor evoked potential (MEP) is an electrical response of peripheral neuro-muscular pathways to stimulation of the motor cortex. MEPs provide objective assessment of electrical conduction through the associated neural pathways, and therefore detect disruption due to a nervous system injury such as spinal cord injury (SCI). In our studies of SCI, we developed a novel, multi-channel set-up for MEP acquisition in rat models. Unlike existing electrophysiological systems for SCI assessment, the set-up allows for multi-channel MEP acquisition from all limbs of rats and enables longitudinal monitoring of injury and treatment for in vivo models of experimental SCI. The article describes the development of the set-up and discusses its capabilities to acquire MEPs in rat models of SCI. We demonstrate its use for MEP acquisition under two types of anesthesia as well as a range of cortical stimulation parameters, identifying parameters yielding consistent and reliable MEPs. To validate our set-up, MEPs were recorded from a group of 10 rats before and after contusive SCI. Upon contusion with moderate severity (12.5mm impact height), MEP amplitude decreased by 91.36±6.03%. A corresponding decline of 93.8±11.4% was seen in the motor behavioral score (BBB), a gold standard in rodent models of SCI.

Connectivity mapping of the human ECoG during a motor task with a time-varying dynamic Bayesian network.

As a partially invasive and clinically obtained neural signal, the electrocorticogram (ECoG) provides a unique opportunity to study cortical processing in humans in vivo. Functional connectivity mapping based on the ECoG signal can provide insight into epileptogenic zones and putative cortical circuits. We describe the first application of time-varying dynamic Bayesian networks (TVDBN) to the ECoG signal for the identification and study of cortical circuits. Connectivity between motor areas as well as between sensory and motor areas preceding and during movement is described. We further apply the connectivity results of the TVDBN to a movement decoder, which achieves a correlation between actual and predicted hand movements of 0.68. This paper presents evidence that the connectivity information discovered with TVDBN is applicable to the design of an ECoG-based brain-machine interface.

Slope analysis of somatosensory evoked potentials in spinal cord injury for detecting contusion injury and focal demyelination.

In spinal cord injury (SCI) research there is a need for reliable measures to determine the extent of injury and assess progress due to natural recovery, drug therapy, surgical intervention or rehabilitation. Somatosensory evoked potentials (SEP) can be used to quantitatively examine the functionality of the ascending sensory pathways in the spinal cord. A reduction of more than 50% in peak amplitude or an increase of more than 10% in latency are threshold indicators of injury. However, in the context of injury, SEP peaks are often obscured by noise. We have developed a new technique to investigate the morphology of the SEP waveform, rather than focusing on a small number of peaks. In this study, we compare SEP signals before and after SCI using two rat models: a contusion injury model and a focal experimental autoimmune encephalomyelitis model. Based on mean slope changes over the signal, we were able to effectively differentiate pre-injury and post-injury SEP values with high levels of sensitivity (83.3%) and specificity (79.2%).

Multiscale entropy analysis of EEG for assessment of post-cardiac arrest neurological recovery under hypothermia in rats.

Neurological complications after cardiac arrest (CA) can be fatal. Although hypothermia has been shown to be beneficial, understanding the mechanism and establishing neurological outcomes remains challenging because effects of CA and hypothermia are not well characterized. This paper aims to analyze EEG (and the alpha-rhythms) using multiscale entropy (MSE) to demonstrate the ability of MSE in tracking changes due to hypothermia and compare MSE during early recovery with long-term neurological examinations. Ten Wistar rats, upon post-CA resuscitation, were randomly subjected to hypothermia (32 degrees C-34 degrees C, N = 5) or normothermia (36.5 degrees C-37.5 degrees C, N = 5). EEG was recorded and analyzed using MSE during seven recovery phases for each experiment: baseline, CA, and five early recovery phases (R1-R5). Postresuscitation neurological examination was performed at 6, 24, 48, and 72 h to obtain neurological deficit scores (NDSs). Results showed MSE to be a sensitive marker of changes in alpha-rhythms. Significant difference (p < 0.05) was found between the MSE for two groups during recovery, suggesting that MSE can successfully reflect temperature modulation. A comparison of short-term MSE and long-term NDS suggested that MSE could be used for predicting favorability of long-term outcome. These experiments point to the role of cortical rhythms in reporting early neurological response to ischemia and therapeutic hypothermia.